Cytogenetic insights into Sosippinae (Araneae, Lycosidae) reveal pronounced diploid number reduction in Aglaoctenus and elevated number of rDNA loci in two unrelated species.

Karyotypes of species sects. Linum and Adenolinum have been studied using C/DAPI-banding, Ag-NOR staining, FISH with 5S and 26S rDNA and RAPD analysis. C/DAPI-banding patterns enabled identification of all homologous chromosome pairs in the studied karyotypes. The revealed high similarity between species L. grandiflorum (2n = 16) and L. decumbens by chromosome and molecular markers proved their close genome relationship and identified the chromosome number in L. decumbens as 2n = 16. The similarity found for C/DAPI-banding patterns between species with the same chromosome numbers corresponds with the results obtained by RAPD-analysis, showing clusterization of 16-, 18- and 30-chromosome species into three separate groups. 5S rDNA and 26S rDNA were co-localized in NOR-chromosome 1 in the genomes of all species investigated. In 30-chromosome species, there were three separate 5S rDNA sites in chromosomes 3, 8 and 13. In 16-chromosome species, a separate 5S rDNA site was also located in chromosome 3, whereas in 18-chromosome species it was found in the long arm of NOR-chromosome 1. Thus, the difference in localization of rDNA sites in species with 2n = 16, 2n = 30 and 2n = 18 confirms taxonomists opinion, who attributed these species to different sects. Linum and Adenolinum, respectively. The obtained results suggest that species with 2n = 16, 2n = 18 and 2n = 30 originated from a 16-chromosome ancestor.

Fish of the genus Selene, known as lookdowns or moonfish, are one of the most morphologically derived groups of the family Carangidae, whose phylogenetic relationships are still largely unknown. In this study, we discuss karyoevolutionary aspects of three representatives of this genus from the Western Atlantic: Selene brownii (2n = 48; FN = 48), Selene setapinnis (2n = 46; FN = 48), and Selene vomer (2n = 48; FN = 50). Their body patterns were also investigated and compared to one another and in relation to two other species of different genera. Two mechanisms of karyotypic evolution seem to have acted in the diversification of this genus, namely pericentric inversions and centric fusions. Mapping of rDNA sequences showed that chromosome pairs bearing 5S rDNA sites are similar, whereas those bearing 18 rDNA sites are morphologically distinct while apparently also exhibiting interspecies synteny. Although the nucleolar organizer-bearing chromosomes are extremely efficient cytotaxonomic markers among Selene species, others cytogenetic patterns of these species are relatively conserved. Hybridization with telomeric probes (TTAGGG)n did not exhibit interstitial telomeric sites (ITS), especially in S. setapinnis, where, along with a reduction in diploid number, a large metacentric pair derived from centric fusion is present. Data obtained by geometric morphometrics enable a clear morphological distinction among the three species, as well as in relation to two other species of the genus Caranx and Oligoplites. Data obtained suggest that morphologic evolution in Selene species was primarily dissociated from visible changes that occurred at the chromosomal level.

Mitotic and meiotic chromosomes of several populations of Eurysternus caribaeus (Coleoptera: Scarabaeidae) were analysed through conventional staining, C-banding, base-specific fluorochromes, silver nitrate staining and fluorescent in situ hybridization (FISH). All specimens showed 2n = 8 in their karyotypes, with a neo-XY sex system (Y is a submetacentric and X a metacentric) and three pairs of submetacentric autosomes. The analysis of constitutive heterochromatin (CH) revealed small blocks located in the centromeric region of all chromosomes which do not present positive staining under the fluorochromes CMA3 and DAPI. Silver nitrate staining revealed that the nucleolar organizer region (NORs) is associated with the sex chromosomes. The FISH technique revealed that rDNA sites in the X and Y are different in size. Data from different populations indicate that the diploid number reduction (2n = 8) observed in E. caribaeus is established and presumably has preceded the dispersion of this species. Moreover, this reduction occasioned the translocation of rDNA sites to the sex chromosomes, X and Y, an uncommon pattern in Scarabaeidae that was observed for the first time by the FISH in this work.

Variability of the 5S and 45S rDNA sites in Passiflora L. species with distinct base chromosome numbers.

Conventional (Giemsa, C-banding, Ag – NORs) and molecular [5S rDNA, 18S rDNA, (TTAGGG)n] cytogenetic techniques were employed to study six species of the genus Eigenmannia Jordan & Evermann, 1896. They exhibited diploid chromosome numbers ranging from 2n=28 (Eigenmannia sp.1) to 2n=38 (Eigenmanniavirescens (Valenciennes, 1836)). The C-banding results revealed that species with the lowest 2n have less heterochromatin content and that morphologically differentiated sex chromosomes observed in two species showed distinct patterns of heterochromatin. While the X1, X2 and Y-chromosomes of Eigenmannia sp.2 showed only centromeric heterochromatin, the XY sex chromosomes of Eigenmanniavirescens possessed large heterochromatic blocks in the terminal position, particularly on the X chromosome. The nucleolus organizer regions (NORs) were located in different positions when compared to the 5S rDNA sites. Additionally, the presence of minor ribosomal gene sites on the sex chromosome pair of Eigenmanniavirescens represented a new type of the sex chromosomes in this group. The telomeric probe (TTAGGG)n hybridized to the terminal portion of all chromosomes in all species examined however, interstitial telomeric sites were found in the metacentric pair No. 2 in Eigenmannia sp.1. The analyzes confirmed some hypotheses about karyotype evolution in the genus Eigenmannia, and brought new information about the distribution of the genetic material in the chromosomes of the samples analyzed providing new insights for understanding the process differentiation in the genomes of species under study.

BackgroundTo gain insight into the evolutionary features of the huntingtin (htt) gene in Chordata, we have sequenced and characterized the full-length htt mRNA in the ascidian Ciona intestinalis, a basal chordate emerging as new invertebrate model organism. Moreover, taking advantage of the availability of genomic and EST sequences, the htt gene structure of a number of chordate species, including the cogeneric ascidian Ciona savignyi, and the vertebrates Xenopus and Gallus was reconstructed.ResultsThe C. intestinalis htt transcript exhibits some peculiar features, such as spliced leader trans-splicing in the 98 nt-long 5' untranslated region (UTR), an alternative splicing in the coding region, eight alternative polyadenylation sites, and no similarities of both 5' and 3'UTRs compared to homologs of the cogeneric C. savignyi. The predicted protein is 2946 amino acids long, shorter than its vertebrate homologs, and lacks the polyQ and the polyP stretches found in the the N-terminal regions of mammalian homologs. The exon-intron organization of the htt gene is almost identical among vertebrates, and significantly conserved between Ciona and vertebrates, allowing us to hypothesize an ancestral chordate gene consisting of at least 40 coding exons.ConclusionDuring chordate diversification, events of gain/loss, sliding, phase changes, and expansion of introns occurred in both vertebrate and ascidian lineages predominantly in the 5'-half of the htt gene, where there is also evidence of lineage-specific evolutionary dynamics in vertebrates. On the contrary, the 3'-half of the gene is highly conserved in all chordates at the level of both gene structure and protein sequence. Between the two Ciona species, a fast evolutionary rate and/or an early divergence time is suggested by the absence of significant similarity between UTRs, protein divergence comparable to that observed between mammals and fishes, and different distribution of repetitive elements.

Duplication and distribution of repetitive elements and non-unique regions in the human genome

BackgroundInformation on the occurrence of sequence features in genomes is crucial to comparative genomics, evolutionary analysis, the analyses of regulatory sequences and the quantitative evaluation of sequences. Computing the frequencies and the occurrences of a pattern in complete genomes is time-consuming.ResultsThe proposed database provides information about sequence features generated by exhaustively computing the sequences of the complete genome. The repetitive elements in the eukaryotic genomes, such as LINEs, SINEs, Alu and LTR, are obtained from Repbase. The database supports various complete genomes including human, yeast, worm, and 128 microbial genomes.ConclusionsThis investigation presents and implements an efficiently computational approach to accumulate the occurrences of the oligonucleotides or patterns in complete genomes. A database is established to maintain the information of the sequence features, including the distributions of oligonucleotide, the gene distribution, the distribution of repetitive elements in genomes and the occurrences of the oligonucleotides. The database can provide more effective and efficient way to access the repetitive features in genomes.

To identify all sesame chromosomes, a multicolour fluorescence in situ hybridization procedure was developed to delineate and physically map the probes of simple sequence repeat (AC)8, 45S rDNA, 5S rDNA, cot-1 DNA and telomeric repeat (TTTAGGG)n. The results showed that the karyotype formula of this species was found to be 2n=6m+18sm+2st. All 13 chromosomes were identified by the banding and colour pattern in the three examined varieties. The FISH analysis of rDNA, (AC)8 and Cot-1 DNA sites showed two sites for 5S rDNA, three sites for 45S rDNA, nine sites for (AC)8 and ten sites for cot-1 DNA. The (AC)8 on the long arm, pericentromeric region of short arm, at the centromeric and near telomeres at the short arm was assigned number 1, 2, 8 and 12, respectively, by simultaneous hybrization with a Cot-1 DNA site. On chromosome 3, we found two (AC)8 sites. One chromosome with no other hybridization signals but a (AC)8 site on the short arm near telomeres was assigned number 11. Both chromosome numbers 5 and 7 had the 45S rDNA and Cot-1 DNA sites near telomeres at the short arm, while the chromosome number 5 was longer than the 7th. The chromosome numbers 6 and 9 had the (AC)8, 5S rDNA and Cot-1 DNA sites near telomeres at the short arm, while the chromosome number 6 was longer than the 9th. The chromosome with the 45S rDNA site at the telomeres and Cot-1 DNA site near the telomeres at the short arm was identified as number 10. The chromosomes with no other hybridization signals but telomeres sites were identified 4 and 13, while the chromosome number 4 was longer than the 13th. It is hoped that the results in this study would provide the reference and support for further studies in genetics and evolution in sesame.

Testicular sperm biopsy combined with intracytoplasmic sperm injection (ICSI) allows men with azoospermia the possibility of fathering a child. However, little information exists on the risk of chromosome abnormalities in their sperm. Multicolor fluorescence in situ hybridization (FISH) analysis was used to determine the frequency of sperm diploidy and disomy for the sex chromosomes in six men with normal karyotypes and non-obstructive azoospermia. A new method using microwave decondensation and codenaturation of sperm nuclei yielded a much larger number of sperm nuclei for FISH analysis than our previous study of men with azoospermia. A total of 59916 sperm were analyzed; more than 9000 sperm were scored for each man. The men with nonobstructive azoospermia had an increased frequency of sperm chromosomal disomy for YY, XY, total sex chromosomal disomy, and diploidy compared with 18 normal controls, but only YY disomy reached statistical significance. One infertile man had a frequency of 3.8% XY disomy and 4.3% diploidy, which was 13-fold and 7-fold higher than control donors, respectively. Our results suggest that some men with nonobstructive azoospermia have a significantly increased frequency of sex chromosomal abnormalities than normal men, but that the overall frequency of abnormalities is similar to that found in infertile men with abnormal semen parameters.

Karyotype analysis of first generation (F1) and backcrossed (BC1) interspecific hybrids were preformatted, suggesting important cytological events in the origin of artificial hybrids. Characterization was performed in 16 backcrossed plants and their genitors by conventional staining, fluorescence in situ hybridization (FISH) with probes for 5S and 45S rDNA and telomeric probes. Genomic identification of hybrids was performed by Genomic In Situ Hybridization (GISH). The chromosome number (2n = 22) was F1 verified in both genitor species P. sublanceolata and P. foetida of the F1 hybrids. However, greater variations in chromosome numbers (2n = 20, 21 and 22) were verified in F1 and BC1 hybrids. A triploid BC1 hybrid was also found suggesting non-reduced female gamete in P. sublanceolata. Variation for number and position of 45S and 5S rDNA sites was also verified in the population, suggesting recombination in homeologous and in non-homeologous chromosomes. Telomeric sites were restricted to terminal regions, discarding the hypotheses of translocation involving telomere sites. GISH clearly distinguished genomes from the genitor species, genome-S (P. sublanceolata) and genome-F (P. foetida), in F1 and BC1 plants, thereby revealing differences in the origin of recombinant chromosomes. Differences in the number and nature of recombinant chromosomes in BC1 hybrids suggests there is high homology between the progenitor genomes, but differences in meiotic recombination in F1 plants. Numerical chromosome modifications, such as chromosome elimination and polyploidization and also chromosome recombination and/or translocations, are hypothesized as common events for the establishment of artificial Passiflora hybrids involving taxa from the Section Dysosmia.

A major goal of genomic and reproductive biology is to understand the evolution of sex determination and sex chromosomes. Species of the 2 genera of the Salamander family Proteidae - Necturus of eastern North America, and Proteus of Southern Europe - have similar-looking karyotypes with the same chromosome number (2n = 38), which differentiates them from all other salamanders. However, Necturus possesses strongly heteromorphic X and Y sex chromosomes that Proteus lacks. Since the heteromorphic sex chromosomes of Necturus were detectable only with C-banding, we hypothesized that we could use C-banding to find sex chromosomes in Proteus. We examined mitotic material from colchicine-treated intestinal epithelium, and meiotic material from testes in specimens of Proteus, representing 3 genetically distinct populations in Slovenia. We compared these results with those from Necturus. We performed FISH to visualize telomeric sequences in meiotic bivalents. Our results provide evidence that Proteus represents an example of sex chromosome turnover in which a Necturus-like Y-chromosome has become permanently translocated to another chromosome converting heteromorphic sex chromosomes to homomorphic sex chromosomes. These results may be key to understanding some unusual aspects of demographics and reproductive biology of Proteus, and are discussed in the context of models of the evolution of sex chromosomes in amphibians.

The diploid number of chromosomes in the male plant is 17 and in the female 16.Of the 17 chromosomes in the male, 14 are autosomes and the remaining 3 sex-chromosomes. The sex-chromosomes are almost equal in size and they are larger than the largest autosomes.There are two V-shaped large X-chromosomes in the somatic division of the female plant, while only one such chromosome beside 2 large J-shaped ones (Y1 and Y2) can be found in the male.The sex chromosomes form a tripartite complex in the reduction division of the pollen mother cell. The X-element in the middle of the tripartite chromosome goes to one pole and the Y1 and Y2 attached to both ends of the X pass to the other.The sex-chromosome complex is not a simple XY pair as first described by WINGE (1923). Such a figure (Fig. 3), from which WINGE drew his conclusion, is only a transitory form to the V-shape arrangement (cf. KIHARA 1927. P. 442). The behavior of the tripartite sex-chromosomes in Humulus japonicus is, therefore, quite identical iwthht atof corresponding stages in Rumex acetcsa.WINGE'S figures (Fig. 11 and 12) representing diakinesis of a pollen mother cell and 16 chromosomes of an archespore mother cell are quite in accordance with my observations.The chromosomal formulae of the plant are as follows:diploid haploid_??_ 14+Y1+X+Y2 7+X, 7+Y1+Y2_??_ 14+X+XThe diploid number of chromosomes in Humulus lupulus (_??_) is 20.

The Heptapteridae family is endemic to the Neotropics and is one of the most representative members of the Siluriformes order in small bodies of water. Due the lack of data for clearly distinguishing their species, only recently was elevated from condition of subfamily to family. However, many doubts about certain species remain yet. There are a relatively large number of cytogenetics studies about heptapterids that permits comparative analyses focused in the karyotype evolution of the group. However, the absence of studies about this theme motivated this work. Were analyzed cytogenetically most representative species of the family: Rhamdia, Imparfinis, Pimelodella, Phenacorhamdia, and Taunaya, and compared with available data from the family. The comparative analysis indicated the predominance of biarmed chromosomes and low variation of the diploid number (around 2n = 58) suggesting that centric inversions were more important than centric fissions in the history of heptapterids. It is corroborated by a remarkable number of NOR phenotypes mainly in Imparfinis and Rhamdia species. A probable condition of Heptapteridae plesiomorphic karyotype would be one composed of 2n = 58 chromosomes of types metacentric and submetacentric mainly, with simple NOR at the sub terminal position. The reduction of diploid number, it would possibly through successive events of chromosome fusion with deletions and inversion rearrangements responsible for NOR variation within the certain genus. The occurrence of B chromosomes is from a derived event, possibly of recent origin, without phylogenetic implications.

Abstract. Donbundit N, Bausriyod P, Tanomtong A, Srisamoot N, Sumontha M, Thongnetr W, Patawang I, Supiwong W, Ditcharoen S, Muanglen N, Kaewmad P. 2022. Karyomorphological delineation, and the NOR loci on the sex chromosome in three species of Chrysopeleinid (Chrysopeleinae: Colubridae) from Thailand. Biodiversitas 23: 3813-3819. The description of chromosomal characteristics of three snakes from Thailand namely, Ahaetulla fusca, A. prasina, and Chrysopelea ornata was by using conventional staining and Ag-NOR banding techniques. The results showed that the same diploid chromosome number (2n) = 36 (16 macrochromosomes + 20 microchromosomes) and the fundamental number (NF) was 54 while the karyotype formulae in these three species are as follows: A. fusca; 2n(36) = Lm6 + Sm2+ Ssm4 + Sa2+ ZZ/ZW + 20 microchromosomes, A. prasina; 2n(36) = Lm6+ Sm6+ Sa2+ ZZ/ZW + 20 microchromosomes and C. ornata; 2n(36) = Lm4+ Lsm2+ Sm2 + Ssm4+ Sa2+ ZZ/ZW + 20 microchromosomes. Besides, the ZZ/ZW sex chromosome was observed in all species, whereas C. ornata was discovered the heteromorphic sex chromosome for the first time. Nucleolar organizer regions (NORs) are located on telomeric region of one pair in all species, while located on microchromosomes in A. fusca and A. prasina, whereas C. ornata was detected on sex chromosomes. Therefore, the Z chromosome revealed the NOR on the telomeric region of the long arm, while NOR of the W chromosome was located on the telomeric region of the short arm.